Reductionist studies of adipose tissue biology require reliable in vitro adipocyte culturing models. Current protocols for induction of adipogenesis in stromal vascular fraction (SVF)-derived preadipocytes require extended culturing periods (12–14d) and have low adipogenic rates. Our objective was to compare the adipogenic efficiency of a 14d-standard induction (STD14) protocol with a 7-d coculture (COC) model using primary mature adipocytes. SVF-derived preadipocytes and mature primary adipocytes were obtained by collagenase digestion from subcutaneous (flank) adipose tissue of dairy cows (n = 6). Primary mature adipocytes were retained for use in COC. SVF was expanded for 2 serial passages and seeded in 24-well plates. Confluent preadipocytes were induced to differentiate with medium containing insulin, troglitazone, 2-isobutyl-1-methylaxanthine (IBMX), dexamethasone, and acetate for 48 h, and then maintained in this medium, excluding the dexamethasone and IBMX, for 7 d in COC, and for 14 d in STD14. Cells on d 7 of induction served as control (STD7). For COC, 0.4-μm transwell inserts with 900 primary mature adipocytes/cm² were placed over the preadipocytes for the first 5 d of adipogenic induction. At the last day of differentiation for each protocol, adipogenic efficiency was evaluated by TAG accumulation (AdipoRed) and gene expression of adipogenesis markers (qPCR). Data were analyzed using One-way ANOVA and Tukey-multiple comparisons. Primary mature adipocytes placed in the inserts in COC were assessed for viability (Vybrant) and decreased 50% from the d 1 to 5 of induction (P = 0.03). Despite this, COC improved adipogenic differentiation of bovine adipocytes as demonstrated by a 3.6 and 3.2 times increase on TAG accumulation compared with STD7 (P = 0.005) and STD14 (P = 0.045), respectively. COC also increased gene expression of peroxisome proliferator-activated receptor-γ (PPARγ, P = 0.0001) and diacylglycerol O-acyltransferase-2 (DGAT2, P = 0.026) compared with STD14. While PPARγ promotes lipid uptake and adipogenesis, DGAT2 enhances TAG synthesis in adipocytes. In conclusion, COC improved adipogenesis efficiency and reduced the time required to induce in vitro differentiation of bovine preadipocytes.